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. 2019 Jul 19;10:3236. doi: 10.1038/s41467-019-11137-5

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — Ab-retargeting after intratumor application of an oncolytic adenovirus leads to improved survival. a Subcutaneous MC38-pSia tumors were established in Ad5-naive mice. After tumor formation, animals received a single application of the oncolytic adenovirus hTert-Ad (by either intratumor or intravenous application) followed by a single i.v. administration of DE1scFv-pSia as shown in the treatment scheme. Control animals were treated by i.t. injection of saline. b Tumor development and survival were monitored (group size n = 6, except control group 0.9 % NaCl: n = 7, median survival: 0.9% NaCl 22 days; hTert-Ad i.v. 23.5 days; hTert-Ad i.t. 27.5 days; hTert-Ad i.v. + DE1scFv-pSia 25 days; hTert-Ad i.t. + DE1scFv-pSia 30.5 days). c Tumor-infiltrating immune cells were analyzed on day 8 after start of i.t. virotherapy (n = 4 in all groups). Proportions of NK cells, macrophages, and MDCSs were calculated as percentage of CD45.2-positive tumor-infiltrating leukocytes. CD8 and CD4 T-cell frequencies were calculated as percentage of CD90.2-positive lymphocytes. d Analyses of tumor antigen-specific CD8 T cells in splenocytes 13 days after i.t. virotherapy (0.9% NaCl: n = 5; hTert-Ad: n = 7; hTert-Ad + DE1scFv-pSia: n = 7). To determine neoantigen-specific responses against Adpgk-R304M, splenocytes were stimulated with the peptide ASMTNMELM, or an irrelevant control peptide, and were analyzed by intracellular staining of IFNγ and flow cytometry. Log-rank (Mantel–Cox) test was used to calculate survival statistics in b. Two-tailed unpaired t test was used to calculate statistics in c and d. *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001. Error bars refer to standard deviation (SD). Source data are provided as a source data file